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目的 探讨核酶在细胞内抑制乙型肝炎病毒基因表达的作用 .方法 计算机设计针对 C基因的三个切点的核酶Rz1,Rz2 ,Rz3,合成核酶基因并将三个核酶基因两两组合克隆入 p GEM7zf(- )质粒中 ,经体外转录后切割靶 RNA;选择Rz2和 Rz3串联的双位点核酶基因构建入逆转录病毒载体p BBS2 12中 ,重组质粒转染 2 .2 .15细胞 ,EL ISA方法分析核酶对乙型肝炎病毒基因表达的抑制作用 .结果 三种不同组合的核酶均可有效切割乙型肝炎病毒基因体外转录底物 .2 .2 .15细胞内表达的双位点核酶可明显抑制 HBe Ag的表达 ,抑制率为 48.6 % ,但对 HBs Ag无明显的抑制作用 .结论 核酶可抑制细胞内乙型肝炎病毒基因的表达 .
Objective To investigate the inhibitory effect of ribozyme on hepatitis B virus (HBV) gene expression in cells.Methods The ribozymes Rz1, Rz2 and Rz3 targeting at the three cleavage sites of C gene were designed and the two ribozyme genes The target RNA was cleaved after in vitro transcription. The double-stranded ribozyme gene Rz2 and Rz3 were selected and inserted into the retroviral vector pBBS2 12, and the recombinant plasmid was transfected into 2.2. 15 cells and EL ISA method to analyze the inhibitory effect of ribozyme on the gene expression of hepatitis B. Results Three different combinations of ribozymes could effectively cleave the in vitro transcription substrate of hepatitis B virus gene.2.2.1.5 Intracellular expression Of double-site ribozyme can significantly inhibit the expression of HBeAg, the inhibition rate was 48.6%, but no significant inhibition of HBsAg.Conclusion The ribozyme can inhibit the expression of hepatitis B virus intracellular genes.